Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of America.
Department of Cellular Physiology and Metabolism, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
PLoS Genet. 2021 Nov 22;17(11):e1009599. doi: 10.1371/journal.pgen.1009599. eCollection 2021 Nov.
microRNAs (miRNAs) are potent regulators of gene expression that function in a variety of developmental and physiological processes by dampening the expression of their target genes at a post-transcriptional level. In many gene regulatory networks (GRNs), miRNAs function in a switch-like manner whereby their expression and activity elicit a transition from one stable pattern of gene expression to a distinct, equally stable pattern required to define a nascent cell fate. While the importance of miRNAs that function in this capacity are clear, we have less of an understanding of the cellular factors and mechanisms that ensure the robustness of this form of regulatory bistability. In a screen to identify suppressors of temporal patterning phenotypes that result from ineffective miRNA-mediated target repression, we identified pqn-59, an ortholog of human UBAP2L, as a novel factor that antagonizes the activities of multiple heterochronic miRNAs. Specifically, we find that depletion of pqn-59 can restore normal development in animals with reduced lin-4 and let-7-family miRNA activity. Importantly, inactivation of pqn-59 is not sufficient to bypass the requirement of these regulatory RNAs within the heterochronic GRN. The pqn-59 gene encodes an abundant, cytoplasmically-localized, unstructured protein that harbors three essential "prion-like" domains. These domains exhibit LLPS properties in vitro and normally function to limit PQN-59 diffusion in the cytoplasm in vivo. Like human UBAP2L, PQN-59's localization becomes highly dynamic during stress conditions where it re-distributes to cytoplasmic stress granules and is important for their formation. Proteomic analysis of PQN-59 complexes from embryonic extracts indicates that PQN-59 and human UBAP2L interact with orthologous cellular components involved in RNA metabolism and promoting protein translation and that PQN-59 additionally interacts with proteins involved in transcription and intracellular transport. Finally, we demonstrate that pqn-59 depletion reduces protein translation and also results in the stabilization of several mature miRNAs (including those involved in temporal patterning). These data suggest that PQN-59 may ensure the bistability of some GRNs that require miRNA functions by promoting miRNA turnover and, like UBAP2L, enhancing protein translation.
microRNAs (miRNAs) 是一种强大的基因表达调控因子,通过在转录后水平抑制靶基因的表达,在多种发育和生理过程中发挥作用。在许多基因调控网络 (GRN) 中,miRNAs 以开关的方式发挥作用,其表达和活性引发从一种稳定的基因表达模式到另一种稳定的、同等稳定的模式的转变,这种模式对于定义新生细胞命运至关重要。虽然具有这种功能的 miRNAs 的重要性是显而易见的,但我们对确保这种形式的调控双稳定性的细胞因子和机制知之甚少。在筛选鉴定导致 miRNA 介导的靶基因抑制无效的时间模式表型的抑制因子的过程中,我们鉴定了 pqn-59,一种人类 UBAP2L 的同源物,作为一种拮抗多种异时性 miRNA 活性的新型因子。具体来说,我们发现 pqn-59 的耗竭可以恢复 lin-4 和 let-7 家族 miRNA 活性降低的动物的正常发育。重要的是,pqn-59 的失活不足以在异时性 GRN 中绕过这些调控 RNA 的要求。pqn-59 基因编码一种丰富的、细胞质定位的、无结构的蛋白,它含有三个必需的“类朊病毒”结构域。这些结构域在体外具有液-液相分离 (LLPS) 特性,正常情况下,它们在体内限制 PQN-59 在细胞质中的扩散。与人类 UBAP2L 一样,在应激条件下,PQN-59 的定位变得非常动态,它重新分布到细胞质应激颗粒中,这对于它们的形成是重要的。来自胚胎提取物的 PQN-59 复合物的蛋白质组学分析表明,PQN-59 和人类 UBAP2L 与涉及 RNA 代谢和促进蛋白质翻译的同源细胞成分相互作用,并且 PQN-59 还与涉及转录和细胞内运输的蛋白质相互作用。最后,我们证明 pqn-59 的耗竭会降低蛋白质翻译,并且还会导致几种成熟 miRNA(包括那些参与时间模式的 miRNA)的稳定。这些数据表明,PQN-59 可能通过促进 miRNA 周转并像 UBAP2L 一样增强蛋白质翻译,来确保需要 miRNA 功能的某些 GRN 的双稳定性。